1. Field of the Invention
This invention relates to an enzyme-substrate system where both the enzyme and substrate are immobilized to solid surfaces and the immobilized enzyme is manipulated such that it can chemically modify the immobilized substrate surface.
2. Description of the Previously Published Art
Many approaches exist for immobilization of enzymes on solid surfaces. Enzymes have been covalently immobilized on chromatography supports for use as a chemically active chromatography column for detecting and/or modifying enzyme substrates passing through the column as described by P. Jadaud et al, J. Chromotography, 476, 165-174 (1989). Alternatively, immobilized enzymes have been used in assays for calorimetric detection of analytes such as the ELISA assay as described by D. J. Reen, Methods in Molecular Biology, 32, 461-6 (1994). As a result of these two technologies, a wide variety of immobilization chemistries have been developed which may be used to maximize the activity of the immobilized enzyme in question. Thus, the art of enzyme immobilization is a mature technique which provides a strong base on which to build this new technology.
In a manner similar to enzyme immobilization, the immobilization of enzyme substrate materials which are capable of reacting with enzymes such as peptides, proteins, lipids, carbohydrates and nucleic acids has been widely studied by many and the following three are just illustrative of that work: Vandenberg et al, J. Colloid Interface Sci, 143, 327-335 (1991); H. Lang et al, Langmuir, 10, 197-210 (1994); and S. Britland et al, Biotechnol. Prog., 8, 155-160 (1992). There have also been several reports which described solution enzyme modification of (solid surface) immobilized substrates as exemplified by T. E. Wilson et al, Langmuir, 10, 1512-1516 (1994) and I. H. Hagestam et al, J. Chromotography, 351, 239-248 (1986). There are no reports, however, indicating that solution phase enzymes have been used to make chemical patterns on substrate surfaces.
Many techniques have been developed for modification and patterning of solid surfaces. These range from conventional lithography utilizing photoresists (including electron and ion beams) as described in the book Semiconductor Lithography by W. Moreau, Plenum Press, New York, (1988); patterning of self-assembled monolayer and silane ultrathin films as described by J. M. Calvert, J. Vac. Sci. Technol., B11, 2155-2163 (1993) and A. Kumar et al, Langmuir, 10, 1498-1511 (1994); and a variety of patterning approaches using scanning probe microscopes as described in The Technology of Proximal Probe Lithography, C. R. K. Marrian, Ed, SPIE Volume IS10, SPEE Press, Bellingham, Wash. (1993). Some of these approaches do exhibit a degree of chemical selectivity (mainly by absorbing photons of specific energy) as seen by the Moreau and Calvert publications, however, none show the spectrum of chemical selectivity possible with enzyme controlled modification. In addition, conventional photoresist patterning requires harsh chemical conditions (organic solvents, elevated temperature) to develop the patterns. Both conventional photoresist and ultrathin film resists require intense photon sources or electron/ion sources to carry out the patterning.
3. Objects of the Invention
It is an object of this invention to provide chemically specific modification of surfaces in a controlled manner.
It is a further object of this invention to provide chemically specific modification of surfaces based on enzyme based catalysis chemistry which uses environmentally benign reaction conditions.
It is a further object of this invention to provide chemical dissection of a surface where the catalytic reactants can be quickly and easily recovered via filtration without fouling the system or the surface.
It is a further object of this invention to provide chemical dissection of a surface by chemical hydrolysis.
It is a further object of this invention to provide chemical modification of a surface where several enzymes could be used on the same substrate to carry out different types of chemically specific modifications.
It is a further object of this invention to provide for a complex series of chemical steps that can be carried on a surface without introduction of harsh solvents or conditions.
It is a further object of this invention to conduct environmentally friendly surface synthesis via enzyme catalyzed chemical reactions.
It is a further object of this invention to provide chemically specific synthesis of surfaces based on enzyme based catalysis chemistry which uses environmentally benign reaction conditions.
It is a further object of this invention to provide chemically-specific patterning and modification of surfaces.
It is a further object of this invention to create complex chemical patterns on a surface.
It is a further object of this invention to provide chemically specific patterning and modification of surfaces based on enzyme based catalysis chemistry which uses environmentally benign reaction conditions.
It is a further object of this invention to create complex chemical patterns on a surface without the use of harsh solvents or conditions.
It is a further object of this invention to create complex chemical patterns on a surface with an enzyme scheme which takes place in an aqueous environment without the need for harsh conditions like organic solvents, high energy beams, or extreme temperature.
It is a further object of this invention to conduct chemical patterning to take place with high chemical selectivity and without ancillary damage to surrounding surface components or the solid substrate material.
It is a further object of this invention to manipulate the position of a single bead coated with immobilized enzymes for high resolution patterning of substrate surfaces.
It is a further object of this invention to place immobilized enzymes on a solid surface which would then be used as a chemical stamp to allow rapid reproduction of a complex pattern on many substrate surfaces.
These and further objects of the invention will become apparent as the description of the invention proceeds.